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LGEUNICAMP

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BIOFUELS

OVERVIEW

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Costs-benefits

• Ideal conditions

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OVERVIEW

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Source: www.bioetanoldecana.org - Adapted

Land use in Brazil

Total land area (851 Mha, 100%)

Agricultural properties area (355 Mha, 42%)

Cropland area (76.7 Mha, 9%)

Sugarcane cropland for energy-use area(3.6 Mha, 0.5%)

OVERVIEW

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Raw Material EnergyProduced/Utilized

Avoided emissions

Sugarcane 9.3 89%

Corn 0.6 – 2.0 -30% to 38%

Wheat 0.97 – 1.11 19% to 47%

Beet 0.97 – 1.11 35% to 56%

Cassava 1.6 – 1.7 63%

Source: www.bioetanoldecana.org

Raw material utilized to produce ethanol

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Source:Statistical Review of world energy 2009 - Adapted

Global ethanol production

OVERVIEW

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Source: D.L. Gazzoni, Adapted

Production and costs ofethanol in Brazil

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ETHANOL

A lot can still be done!

OVERVIEW

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Good substitutes for oil-based fuels

Solve the environmental problems

OVERVIEW

Biofuels

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Biofuels do not compete with food and land use

OVERVIEW

Biofuels versus Food

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Enable appropriate land use

OVERVIEW

Scientific research

Should continue - obtain more efficient biofuels using genetic

engineering

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ETHANOL

Project

Costs

Land use

OVERVIEW

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OVERVIEW

How is ETHANOL produced in Brazil?

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CONTAMINANTS

OVERVIEW

How is ETHANOL produced in Brazil?

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iGEM project - CONTAMINATION

YEAST

Ethanol Insulin, yogurt

BACTERIA

OVERVIEW

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… Prepare

yourselves to get into the

microguards´lives

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RECOGNITION SYSTEM

KILLING SYSTEM

The YeastguardOVERVIEW

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First Mechanism (increased lactate sensibility)

Second Mechanism (recognizing lactate on the interior of the cell)

Biobrick submitted! Biobrick submitted!

Recognitionsystem

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RECOGNITION SYSTEM

KILLING SYSTEM

The YeastguardOVERVIEW

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Biobrick submitted!

killingsystem

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RECOGNITION SYSTEM

KILLING SYSTEM

YeastguardOVERVIEW

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Successful assembled biobrick

BBa_K284002: JEN1 Promoter

BBa_K284003: Partial DLD promoter

BBa_K284023: EYFP regulated by ADH1 promoter (characterization device)

BBa_K284001: Lysozyme from Gallus gallus

BBa_K284016: Lysozyme constitutive expression (characterization device)

BBa_K284017: Lysozyme under control of DLD promoter (characterization device)

THEYeastguARdresults

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Recognition system

Differentiation system

Killing system

The coliguardOVERVIEW

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Briefly,

If contaminants conjugate with our coliguard:

1. Induces Py

2. Releases AI2

3. Activate Differentiation and Killing mechanism

If contaminants release AI2 themselves:

1. .

2. .

3. Activate Differentiation and Killing mechanism

skip steps!

Recognitionsystem

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Recognition system


Killing system


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Worker Cells Killer Cells

Amount Amount

Basal proportion!

Differentiationsystem

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Absence of contaminants:


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Presence of contaminants:


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Presence of contaminants:

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Elimination of contaminants:

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Absence of contaminants:


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Worker lineage characteristics!

The slippage mechanism controlling the basal proportions

(AGTC)10


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The slippage mechanism controlling the basal proportions

(AGTC)10(AGTC)9

Killer lineage characteristics!


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Integration between the mechanisms of slippage and Cre-Recombinase

Slippage error doesn’t occurs (most cases):

Cell Cycle!

Conjugation Inhibition System

Worker lineage


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Killer lineage

Integration between the mechanisms of slippage and Cre-Recombinase

Slippage error occurs (few cases):

(AGTC)9


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Enhanced Amount of Killers cells induced by contaminants


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Recognition system


Killing system


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Killingsystem

The Kamikaze System

The Colicin System

CeaB lethal gene (colicin E2) into F plasmid

CeiB antidote gene into genomic DNA

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Recognition system


Killing system


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Successful assembled biobrick

BBa_K284008: Py promoter + RFP device (characterization device)

BBa_K284031: Cre-Recombinase without ATG start codon

BBa_K284022: T4 endolysin under control of T7 promoter (characterization device)

BBa_K284022

THE COLIGUARD REsults

Characterized parts

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The coliguardresults

Characterizing BBa_K284022

Transformations into E. coli strain C43 (T7 promoter is induced by IPTG)

Grown inocula were diluted to starter OD=0,2

Once OD=0,8 was reached induction with IPTG

Incubation for 4 hours at 37⁰C

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Plated each culture into solid LB-AMP media


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SDS-PAGE


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Summary

2 contamination control models designed, one for prokaryotes and one for eukaryotes

30 new biobrick parts and devices designed

9 new biobrick parts and devices constructed and submitted to the registry

4 new biobrick parts or devices were tested

1 new biobrick device worked as expected (BBa_K284022)

An existing Biobrick part was characterized (BBa_K112806)

A new approach to an issue of Human Practice in synthetic biology as it relates to our project was outlined and detailed

Accomplishments

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This work could not be done without the sponsor of

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Special Acknowledgements:

Prof. Dr. Fernando Costa. Dean, UNICAMP.

Prof. Dr. Luís Cortez. CORI, UNICAMP.

Prof. Dr. Fábio Papes. Dept. Genética, Evolução e Bioagentes, UNICAMP.

Prof. Dr. Paulo Arruda. Dept. Genética, Evolução e Bioagentes, UNICAMP.

Prof. Dr. Gonçalo A. G. Pereira. Dept. Genética, Evolução e Bioagentes, UNICAMP.

Support:

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thank you!!!!

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